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. 2023 Jul 26;28(15):5666.
doi: 10.3390/molecules28155666.

Quality Control and Authentication of Black Betel Leaf Extract (Piper acre Blume) from East Kalimantan as an Antimicrobial Agent Using a Combination of High-Performance Liquid Chromatography and Chemometric Fourier Transform Infrared

Vina Maulidya  1   2 , Aliya Nur Hasanah  1 , Laode Rijai  2 , Muchtaridi Muchtaridi  1   3
Affiliations

Quality Control and Authentication of Black Betel Leaf Extract (Piper acre Blume) from East Kalimantan as an Antimicrobial Agent Using a Combination of High-Performance Liquid Chromatography and Chemometric Fourier Transform Infrared

Vina Maulidya et al. Molecules. .

Abstract

Black betel leaf from East Kalimantan contains various secondary metabolites such as alkaloid saponins, flavonoids, and tannins. A compound, piperenamide A, which has antimicrobial activity, is also found in black betel leaf. This study aims to identify and authenticate the compound piperenamide A found in black betel leaf extract in other types of betel plant using HPLC and FTIR-chemometrics. The extraction method used was maceration with 70% ethanol solvent. Determination of piperenamide A content in black betel leaf extract was via HPLC column C18, with a maximum wavelength of 259 nm and a mobile phase of water:acetonitrile at a flow rate of 1 mL/minute. From the results, piperenamide A was only found in black betel (Piper acre) and not in Piper betel and Piper crocatum. Piperenamide A levels obtained were 4.03, 6.84, 5.35, 13.85, and 2.15%, respectively, in the samples studied. The combination of FTIR spectra with chemometric methods such as PCA and PLS-DA was used to distinguish the three types of betel. Discriminant analysis can classify black betel (Piper acre), Piper betel, and Piper crocatum according to its type. These methods can be used for identification and authentication of black betel.

Keywords: FTIR; HPLC; PCA; PLS-DA; black betel; ethanol extract; piperenamide A.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Structure of the piperenamide A compound [4].
Figure 2
Figure 2
Types of betel plants: (a) Piper acre Blume; (b) Piper betle L.; and (c) Piper crocatum.
Figure 3
Figure 3
Antimicrobial activity of Piper betle var. nigra extract. (a) Antibacterial activity against Streptococcus mutans; and (b) antibacterial activity against Streptococcus sanguinis. 1 (concentration of 1%); 2 (concentration of 0.5%) [4].
Figure 4
Figure 4
Chromatographic separation of (a) standard piperenamide A; (b) samples of black betel extract; (c) Piper betel extract samples; and (d) samples of red betel extract using a Purospher RP−18 column under a gradient elution of water and acetonitrile.
Figure 5
Figure 5
FTIR results of standard piperenamide A and the five samples. (a) FTIR spectra of piperenamide A; (b) FTIR spectra of samples of black betel leaf extract, green betel leaf extract, and red betel leaf extract; and (c) FTIR spectra of 5 samples of black betel extract from East Kalimantan.
Figure 6
Figure 6
Overview of PCA chemometric results using MetaboAnalyst software 5.0. Annotation: purple circle: Piper betle sample; blue circle: Black betle sample; red circle: Red betle sample.
Figure 7
Figure 7
Overview of PLS−DA chemometric results using MetaboAnalyst software 5.0.
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